Selection in Agrobacterium-mediated transformation Which selectable marker enzymes are routinely used to recover plant cells transformed by Agrobacterium?

Difficulty: Easy

Correct Answer: Any of the above, depending on the vector and plant species

Explanation:


Introduction / Context:
Selectable markers enable researchers to isolate the relatively few plant cells that acquire and express T-DNA after Agrobacterium infection. Understanding common marker enzymes and antibiotics is fundamental for designing transformation experiments and interpreting regeneration results.


Given Data / Assumptions:

  • Agrobacterium delivers T-DNA harboring a selectable marker gene into plant cells.
  • Markers typically confer antibiotic resistance that permits growth on selective media.
  • Different crops and protocols favor different antibiotic/marker combinations.


Concept / Approach:
Widely used markers include neomycin phosphotransferase II (NPTII; kanamycin/G418 resistance) and hygromycin phosphotransferase (HPT; hygromycin resistance). Streptomycin/spectinomycin resistance cassettes are also used in some systems. The best choice depends on plant sensitivity, background tissue responses, and regulatory considerations.


Step-by-Step Solution:

List common markers: NPTII (kanamycin), HPT (hygromycin), and other aminoglycoside phosphotransferases.Match to selection media: tissues survive only if the marker is expressed.Note species- and tissue-specific sensitivities to antibiotics.Conclude that any of the listed marker enzymes may be used, depending on context.


Verification / Alternative check:
Transformation protocols across tobacco, tomato, rice, and Arabidopsis document successful selections using kanamycin or hygromycin; some use streptomycin/spectinomycin in plastid or nuclear selection schemes.


Why Other Options Are Wrong:

  • NPTII or HPT alone: each is valid but not exclusive.
  • Streptomycin phosphotransferase alone: valid in some cases but not the only standard.
  • lacZ alone: a reporter, not a robust selectable marker in plants.


Common Pitfalls:
Using antibiotic levels that either fail to suppress escapes (too low) or kill regenerable transformants (too high). Always empirically determine minimal lethal concentrations for the target tissue.


Final Answer:
Any of the above, depending on the vector and plant species

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